A review of the use of genetically engineered enzymes in electrochemical biosensors.
ABSTRACT This article gives an overview of the electrochemical biosensors that incorporate genetically modified enzymes. Firstly, the improvements on the sensitivity and selectivity of biosensors that integrate mutated enzymes are summarised. Next, new trends focused on the oriented immobilisation of mutated enzymes through specific functional groups located at their surface are reviewed. Finally, the effect of enzyme mutations on the electron transfer distance and kinetics of electrochemical biosensors is described.
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ABSTRACT: This work presents an application of automatic flow based biosensor to detect binary (chlorpyriphos-oxon (CPO) and malaoxon (MO)) organophosphate (OP) mixtures in milk, based on artificial neural network (ANN). Genetically modified acetylcholinesterase (AChE) B394 and B4 were used as a biological recognition element for sensor development. AChE binds with OPs irreversibly, creating an anionic phosphonyl species. The enzymes were coupled on screen printed electrodes (SPEs) and inserted in a flow cell connected to the potentiostat and syringe pump. In order to model the combined response of CPO and MO, a total set of 19 mixtures were prepared using ANN. The modeling was validated with an external test of 6 milk samples spiked with CPO and MO mixtures. The spiked concentrations of CPO and MO were ranged from 5 × 10−10 to 5 × 10−12 M and 1.01 × 10−10 to 9.17 × 10−11 M, respectively. These concentrations were determined using factorial designing (FD) method and the obtained and expected recovery values in milk showed good co-relation. The average % recovery yields for CPO and MO are 109.53 and 100.66, respectively.Sensors and Actuators B Chemical 03/2015; 208. DOI:10.1016/j.snb.2014.11.011 · 3.84 Impact Factor
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ABSTRACT: In recent year’s novel bioinformatics tools have been developed to design, rationalize, and optimize the classical techniques to covalently attach proteins to solid surfaces. In this sense, the rational design of immobilized derivative (RDID) has become a potent tool to predict the orientation of proteins covalently attached to the support, the reactivity of protein interacting groups, the maximum protein quantity to immobilize, as well as the protein regions involved in adsorption to the support. RDID include an array of algorithms implemented in the computer program RDID1.0 which take into account the properties of the ligand (3D structure, superficial arrangement of protein interacting groups and its reactivity, etc.) as well as the textural properties of the support during the predictions. In silico analyses may help to establish the optimal immobilization conditions and to understand the behavior of immobilized enzymes. RDID application allows the development of optimal immobilized systems with a great performance in enzymatic bioconversion processes. Novel application of RDID strategy to acylase immobilized biocatalysts and site-directed mutagenesis are described in this work.Breaking news in bioinformatics applied to covalent immobilization of acylases biocatalyst, Simposio Internacional de Química 2013 (SIQ´13), Cayo Santa Maria, Villa Clara, Cuba.; 06/2013
101 edited by Paul C Guest and Sabine Bahn, 11/2011; Academic Press., ISBN: 0123877180